More than 80% of global transmission of HIV infection occurs via mucosal routes. The ability of vaccines to induce mucosal immunity may be required for protection against HIV infection and the immunodeficiency syndrome that emerges after infection. Various AIDS vaccine candidates are currently under development, each with potential advantages and disadvantages. DNA vaccines have features that make them attractive vaccine candidates for HIV and the combination of DNA vaccination plus recombinant vaccinia is one of the favored approaches in AIDS vaccine development. We have established a vaccination regimen via the nasal route that stimulates mucosal and systemic responses with a SHIV DNA vaccine, which consists of a single plasmid carrying a SHIV proviral genome that produces noninfectious particles. We have the appropriate tools to measure SHIV- and SlV-specific IgA levels in mucosal secretions, mucosal cell-mediated responses, and SHIV and SlV-specific humoral and cellular systemic responses. As a consequence, we believe that we are well placed to establish the effectiveness of mucosal vaccination and how stimulation of virus-specific mucosal and systemic responses may affect the efficacy of a HIV/SIV vaccine. This proposal is designed to extend on going studies by investigating approaches restricted to the nasal route and evaluate their efficacy in female animals. Towards this goal we propose: 1) to evaluate whether the addition of different cytokines to the DNA priming portion of the SIV DNA-MVA regimen leads to both systemic and mucosal virus specific immune responses that are different in magnitude or in mucosal/systemic ratio than those observed with SHIV-IL-2/Ig; 2) to compare which of two immunization regimens (SIV+ cytokines DNA priming followed by boosting with 1. recombinant adenovirus or 2. genitically inactivated SIV particles) administered via the nasal route better engenders high levels of both mucosal and systemic immune responses in female macaques that can provide significant control of viremia and disease progression. The results of this investigation may have important implications for AIDS vaccine development and for the development of DNA vaccines for other mucosal pathogens.